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Biomedical subjects

M Yamaguchi

Publications and source records attributed to M Yamaguchi.

At least 775 records · Page 43Linked to original sources

Increase of (Ca(2+)-Mg2+)-ATPase activity in hepatic plasma membranes of rats administered orally calcium: the endogenous role of regucalcin.

The alteration of the plasma membrane (Ca(2+)-Mg2+)-ATPase activity in the liver of rats administered orally calcium chloride solution was investigated. The plasma membrane (Ca(2+)-Mg2+)-ATPase activity was significantly increased by a single oral administration of calcium (10, 25 and 50 mg/100 g body weight) in rats. This increase was seen between 10 and 60 min after the administration. The presence of anti-regucalcin IgG (1.0-5.0 micrograms/ml) in the enzyme reaction mixture caused a complete inhibition for the elevation of the plasma membrane (Ca(2+)-Mg2+)-ATPase activity by the addition of regucalcin (0.25 microM). Also, the calcium administration-induced increase in hepatic plasma membrane (Ca(2+)-Mg2+)-ATPase activity was completely abolished by the presence of anti-regucalcin IgG (1.0 and 2.5 micrograms/ml). Moreover, the calcium administration-induced increase in hepatic plasma membrane (Ca(2+)-Mg2+)-ATPase activity was not inhibited by vanadate (0.1 and 0.2 mM) addition into the enzyme reaction mixture, although the inhibitory effect of vanadate was seen in the plasma membranes from normal rat liver. Now, the activating effect of regucalcin (0.25 microM) on hepatic plasma membrane (Ca(2+)-Mg2+)-ATPase was not inhibited by vanadate addition. The endogenous regucalcin may play a role in the calcium administration-induced increase of (Ca(2+)-Mg2+)-ATPase activity in the liver plasma membranes of rats.

Animals↗

17 beta-Estradiol stimulates the expression of hepatic calcium-binding protein regucalcin mRNA in rats.

The effect of nuclear receptor-related hormones on the expression of hepatic calcium-binding protein regucalcin mRNA in rats was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb of open-reading frame). A single subcutaneons administration of 17 beta-estradiol (0.5, 1.0 and 2.0 mg/kg body weight) in rats induced a remarkable increase of regucalcin mRNA in liver; the level was about 200% of control at 24 h after the administration of 2.0 mg/kg. The increase showed about 350% even at 6 h after the administration. Meanwhile, hepatic regucalcin mRNA level was not appreciably altered by a single subcutaneous administration of thyroxine (T4) (20, 40 and 80 mg/kg) or hydrocortisone (10 and 30 mg/kg) in rats. The present study demonstrates that the expression of hepatic regucalcin mRNA is stimulated by estrogen action in the liver nuclei of rats.

Animals↗

Calcium administration increases calcium-binding protein regucalcin concentration in the liver of rats.

The alteration of regucalcin concentrations in the liver and serum of rats administered orally calcium is investigated. Rats received a single oral administration of calcium chloride solution (25, 50 and 75 mg Ca/100 g body weight). The administration of calcium (50 mg/100 g) produced a significant increase in liver regucalcin concentration between 30 and 180 min after the administration, while serum regucalcin concentration was not altered appreciably. The effect of calcium administration increasing liver regucalcin concentration was also seen with the dose of 25 mg/100 g. When liver cytosol prepared from normal rats was incubated for 6 h in the presence of 10 microM Ca2+, the cytosolic regucalcin concentration at 3 and 6 h of incubation was decreased about 20% (p < 0.05) as compared with the value at zero time point, indicating that the presence of Ca2+ does not inhibit the decomposition of liver cytosolic regucalcin. Moreover, serum regucalcin concentration was not significantly altered by the incubation for 6 h at 37 degrees C, indicating a stability of regucalcin in rat serum. This suggests that the calcium administration-induced in liver regucalcin concentration is not based on the inhibition of regucalcin release from liver to serum. The present study demonstrates that regucalcin in the liver is clearly increased by calcium administration, presumably due to stimulating the protein synthesis.

Animals↗

Specific species and tissue differences for the gene expression of calcium-binding protein regucalcin.

The existence and expression of gene encoding the Ca(2+)-binding protein regucalcin in various species and tissues were investigated with Southern and Northern hybridization analyses using regucalcin cDNA (0.9 kb of open reading frame). Genomic Southern hybridization analysis demonstrated that regucalcin gene was widely conserved among higher animals including human, monkey, rat, mouse, dog, bovine, rabbit and chicken. The gene was not found in yeast. The Northern blot analysis of poly (A) +RNAs extracted from the liver of various species showed that regucalcin mRNA was predominantly expressed in rat and mouse, although the expression was also seen in human, bovine and chicken. Furthermore, the enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG indicated that hepatic regucalcin concentration was most pronounced in rat as compared with that of guinea pig, mouse and chicken. These observations show that the gene expression of regucalcin and its protein synthesis is unique in the liver of rats, suggesting the existence of a specific mechanism in demonstrating regucalcin synthesis from gene.

Animals↗

Chemotactic agonists induce cytokine generation in eosinophils.

Recent studies have shown that eosinophils are capable of generating and releasing cytokines, providing a novel biologic aspect of eosinophils for regulating allergic inflammation by an autocrine or paracrine mechanism. Eosinophils synthesize various cytokines; however, the physiologic stimuli that trigger eosinophils to generate cytokines have not been fully elucidated. We examined the effect of chemotactic agonists on eosinophil cytokine generation by employing the determination of IL-8 as the main parameter. Both C5a and FMLP stimulated eosinophils to release IL-8, whereas platelet-activating factor and C-C chemokines did not exert any significant effects. On a molar basis, C5a was two orders of magnitude more potent than FMLP. The generation of IL-8 by chemoattractants was absolutely dependent on the presence of cytochalasin B. Pertussis toxin completely attenuated C5a- and FMLP-induced IL-8 production, indicating the involvement of pertussis toxin-sensitive G-proteins in the signal-transduction process leading to these responses. Experiments of in situ hybridization and PCR amplification revealed that both C5a and FMLP promoted eosinophil IL-8 production through transcriptional gene activation. Pyrrolidine dithiocarbamate completely abrogated chemoattractant-induced IL-8 production, indicating the involvement of NF-kappa B in the cytoplasmic/nuclear signal-transduction process. Furthermore, chemoattractant-induced cytokine production was not limited to IL-8; C5a and FMLP but not platelet-activating factor induced significant secretion of granulocyte-macrophage-CSF from eosinophils. These results indicate that C5a and FMLP stimulate eosinophils to elaborate cytokines, which could be an important mechanism in the regulation of allergic inflammation.

Base Sequence↗

Expression of hepatic calcium-binding protein regucalcin mRNA is elevated by refeeding of fasted rats: involvement of glucose, insulin and calcium as stimulating factors.

The effect of refeeding on the expression of Ca(2+)-binding protein regucalcin mRNA in the liver of fasted rats was investigated. When rats were fasted overnight, the hepatic regucalcin mRNA level was reduced about 70% of that in feeding rats. Refeeding produced a remarkable elevation of hepatic regucalcin mRNA level (about 150-170% of fasted rats). Liver regucalcin concentration was appreciably increased by refeeding, although it was not altered by fasting. The oral administration of glucose (2 g/kg body weight) to fasted rats caused a significant increase in hepatic regucalcin mRNA level. Moreover, hepatic regucalcin mRNA level was clearly elevated by a single subcutaneous administration of insulin (10 and 100 U/kg) to fasted rats. The hormonal effect was not further enhanced by the simultaneous administration of calcium chloride (250 mg Ca/kg) to fasted rats, although calcium administration stimulated regucalcin mRNA expression in the liver. The present study suggests that the expression of hepatic regucalcin mRNA stimulated by refeeding is significantly involved in the action of insulin and/or calcium as stimulating factors.

Animals↗

Proliferation and differentiation of myelodysplastic CD34+ cells: phenotypic subpopulations of marrow CD34+ cells.

In a search for a mechanism to explain the impaired growth of progenitor cells in patients with myelodysplastic syndromes (MDS), marrow CD34+ cells were purified up to 94.9% +/- 4.2% for normal individuals and 88.1% +/- 17.6% for MDS patients, using monoclonal antibodies and immunomagnetic microspheres (MDS CD34+ cells). Phenotypic subpopulations of these CD34+ cells were analyzed for CD38, HLA-DR, CD33, CD13, CD14, CD41 and CD3 plus CD19, in association with proliferative and differentiative capacities. The 15 studies performed included 12 MDS patients. Coexpression rate of CD13 significantly increased in the MDS CD34+ cell population with a value of 91.4% +/- 11.6% and ranging from 60.3% to 100%, and exceeded 99% in four studies, whereas that of normal CD34+ cells was 49.9% +/- 15.8%, ranging from 28.2% to 70.1% (P < .001). Coexpression rate of CD38, HLA-DR, CD33, CD14, and CD3 plus CD19 in MDS CD34+ cells did not significantly differ from that of normal CD34+ cells. The total number of colonies and clusters grown from 100 normal marrow CD34+ cells was 40.4 +/- 8.6, the range being from 27.2 to 50.3; this varied in MDS marrow CD34+ cells with a value of 34.0 +/- 28.7, the range being 0 to 95.9. The lineage of colonies and clusters promoted by MDS marrow CD34+ cells was predominantly committed to nonerythroid with impaired differentiation in 13 of 15 studies (87%). CD13 is first expressed during hematopoiesis by colony-forming unit granulocyte-macrophage and is absent in erythroid progenitors. Therefore, this study provides direct evidence for the lineage commitment of MDS CD34+ cells to nonerythroid with impaired differentiation and explains the mechanism of nil or low colony expression of MDS progenitor cells to erythroid lineage.

Aged↗

Adult T-cell leukemia/lymphoma with two distinct clones in the peripheral blood and lymph node.

A case of adult T-cell leukemia/lymphoma (ATL) with two different clones in the peripheral blood and lymph nodes is reported here. When cellular DNA from the lymph node was digested with EcoRI, one band larger than 9 Kb was detected. Digestion of the cellular DNA with PstI resulted in one clear band in addition to three internal fragments. In contrast, when cellular DNA from malignant peripheral blood lymphocytes (PBL) was digested with the same endonucleases, distinct bands at positions different from those observed in the lymph node were detected, indicating two separate malignant clones in the patient. Monoclonality of the tumor cells was shown by T-cell receptor-beta (TCR-beta) gene rearrangement in both PBL and lymph node. Furthermore, there was a difference in the surface phenotype between tumor cells taken from peripheral blood (CD4+, CD8-) and lymph node (CD4+, CD8+). These findings suggest the presence of two different ATL clones in PBL and lymph node in a single patient simultaneously, which is distinguishable by the integration pattern of human T-cell leukemia virus type I (HTLV-I) proviral DNA.

Aged↗

Enhanced expression of calcium-binding protein regucalcin mRNA in regenerating rat liver.

The expression of hepatic calcium-binding protein regucalcin mRNA was investigated in regenerating rat liver. The change of regucalcin mRNA levels was analyzed by Northern blotting, using liver regucalcin cDNA (0.9 kb with complete open reading frame). The reduced liver weight by partial hepatectomy (about 70%) was completely restored at 3 days after surgery. Regenerating liver significantly increased calcium content. Liver regucalcin mRNA levels clearly increased 1-5 days after hepatectomy, in comparison with that of sham-operated rats, although the increase was not seen 12 hr after the surgery. Increased regucalcin mRNA levels in regenerating liver were appreciably reduced by single intraperitoneal administration of actinomycin D (100 micrograms/100 g body weight), an inhibitor of transcriptional process. Moreover, the increased regucalcin mRNA levels by hepatectomy was weakened by a single intraperitoneal administration of trifluoperazine (2.5 mg/100 g), an inhibitor of Ca2+/calmodulin. These findings demonstrate that the expression of hepatic regucalcin mRNA is enhanced in regenerating rat liver.

Analysis of Variance↗

DNase I interaction on muscle Z-line.

The effect of deoxyribonuclease I on muscle Z-line structures was re-examined. Under conditions of deoxyribonuclease I activation (presence of the divalent cation Ca2+ and Mg2+), a deoxyribonuclease I preparation did not affect Z-line structure if phenylmethylsulfonylfluoride, an inhibitor of serine proteases, was also present. In the absence of protease inhibitor, both Z-lines and M-lines were digested, even in the presence of EDTA and EGTA as inhibitors of deoxyribonuclease I. These electron microscopic observations were consistent with the following results from sodium dodecyl sulphate gel electrophoresis: when the protease was inhibited but deoxyribonuclease I was activated, myofibrillar proteins remained essentially intact. However, degradation of proteins in both rabbit psoas and chicken pectoralis myofibrils was observed in the presence of deoxyribonuclease I inhibitors when the protease inhibitor was absent. Our data strongly suggest that the interaction of deoxyribonuclease I with Z-line proteins previously reported is most likely due to contamination of the deoxyribonuclease I fraction by the serine-type proteases.

Animals↗

Effects of adenosine diphosphate on the structure of myosin cross-bridges: an X-ray diffraction study on a single skinned frog muscle fibre.

Using a technique to obtain a detailed X-ray diffraction pattern from a single skinned frog muscle fibre, we studied the effects of ADP on the structure and arrangement of myosin heads. An imaging plate and a cooled-CCD X-ray detector were used to record the diffraction patterns. Addition of 1 mM ADP to a rigor fibre increased the intensity of the third-order meridional reflection of the myosin repeat by 50-85%. The intensity of the sixth-order meridional reflection also increased. After removing the ADP, these intensities decreased but did not return to the level before the ADP was added. No significant changes were observed in the intensities of the equatorial reflections and the actin layer-lines. These results suggest that, upon ADP binding, the conformation of a myosin head changes without detaching from actin. The structural change may involve a relative motion between domains of the myosin head by the closure of the cleft to which an ADP molecule binds.

Adenosine Diphosphate↗

Zinc decrease and bone metabolism in the femoral-metaphyseal tissues of rats with skeletal unloading.

Whether the decrease of zinc content in the femoral-metaphyseal tissues of rats with skeletal unloading is involved in the alteration of bone metabolism was investigated. Skeletal unloading was designed using the model of hindlimb suspension in rats. Animals were fed for 4 days with the unloading. The metaphyseal zinc content were significantly decreased by the unloading. Zinc accumulation in the metaphyseal tissues by a single oral administration of zinc sulfate (20 mg Zn/100 g body weight) was partially depressed by the unloading, although serum zinc concentration was higher than that in normal rats, suggesting an impaired movement of zinc from serum into bone tissues by the unloading. Skeletal unloading caused a significant decrease of alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the metaphyseal tissues. These decreases were completely restored by addition of zinc sulfate (10(-4) M) or beta-alanyl-L-histidinato zinc (AHZ; 10(-5) M) in a culture medium with the metaphyseal tissues in vitro. The effects of zinc compounds were abolished by the presence of cycloheximide (10(-8) M), suggesting that the zinc effect is based on a newly synthesized protein. Dipicolinate (10(-4) and 10(-5) M), a potent zinc-chelating agent, caused an appreciable decrease of zinc content and alkaline phosphatase activity in the metaphyseal tissues. This decrease was restored by zinc supplement. The present results suggest that the skeletal unloading-induced decrease of zinc content in the femoral-metaphyseal tissues plays a role in the deterioration of bone metabolism in the unloaded rats.

Alkaline Phosphatase↗

Mantle cell lymphoma with the features of mucosa-associated lymphoid tissue (MALT) lymphoma in an HTLV-I-seropositive patient.

A case of small lymphocytic B-cell lymphoma with seropositivity for human T-cell leukemia virus type I (HTLV-I), whose clinical features were closely related to those of mucosa-associated lymphoid tissue (MALT) lymphoma, is presented. The neoplastic cells of the lymph node were immunologically positive for CD5, in addition to several B-cell markers, but negative for CD10, and cytogenetically carried a t(11;14)(q13;q32). These findings were fully consistent with so-called mantle cell lymphoma (MCL). In addition to the lymph nodes and bone marrow, multiple extranodal sites including lacrimal and salivary glands, lung and stomach (where MALT is present) were occupied by lymphoma cells. These extranodal lesions were immunologically identical to the lymph nodes (CD5(+), CD10(-)), but histologically showed lymphoepithelial lesions (LEL) characteristic of MALT lymphoma. These findings suggest a possible relationship between MCL and MALT lymphoma, and the neoplastic cells are thought to originate from the CD5-positive B cells, which are present near the areas across the mantle and marginal zones. Furthermore, HTLV-I-infection, which appears to create an immunodeficient state or modulate the B-cell response, is thought to play a role in B-cell lymphomagenesis.

Antigens, CD↗

Risk factors for the development of acute disseminated intravascular coagulation in patients with systemic lupus erythematosus.

Since acute disseminated intravascular coagulation (DIC) often contributes to a fatal outcome in patients with systemic lupus erythematosus (SLE), prediction of its development is important to prevent the occurrence of such an event. To analyze the risk factor(s) contributing to the development of acute DIC in SLE, we carried out a retrospective study of a series of 129 SLE patients, eight of whom developed DIC during the course of this disease, to assess which of the easily assessable parameters, present at the time of first medical examination, were of predictive significance. The important individual variables, determined by univariate analysis, were male sex, leukopenia, and infection. These factors were placed in a multivariate logistic regression model, and only one factor, infection at first medical examination, was found to have predictive significance for the development of acute DIC in SLE patients. The prevention and control of infection in SLE patients might have implications for preventing the development of acute DIC.

Acute Disease↗

Endothelin-1 and big endothelin-1 increase in human endometrium during menstruation.

OBJECTIVES: Although the physiologic and pathologic roles of endothelin-1 in reproduction have been investigated, little is known about human uterine tissue levels. We studied the levels of immunoreactive endothelin-1 and immunoreactive big endothelin-1 in human endometrium and myometrium during each menstrual phase. STUDY DESIGN: Materials were obtained at hysterectomy (endometrium, n = 33; myometrium, n = 27). We measured immunoreactive endothelin-1 and immunoreactive big endothelin-1 by radioimmunoassay and performed an immunohistochemical study of the tissue. Data were analyzed by the Mann-Whitney U test. RESULTS: We detected larger amounts of immunoreactive endothelin-1 and immunoreactive big endothelin-1 in the endometrium than in the myometrium throughout the menstrual, proliferative, and secretory phases. Endometrial immunoreactive endothelin-1 and immunoreactive endothelin-1 were significantly increased in the menstrual phase (endothelin-1 68.8 +/- 23.3 pg/mg protein, n = 5, p < 0.005; big endothelin-1 45.2 +/- 5.7 pg/mg protein, n = 5, p < 0.003) compared with the other phases (endothelin-1 30.7 +/- 9.5 and 30.5 +/- 14.0 pg/mg protein; big endothelin-1 19.9 +/- 6.7 and 24.1 +/- 7.4 pg/mg protein). Immunohistochemistry demonstrated that the endometrial stromal cells were positive for antiendothelin monoclonal antibody only in the premenstrual and menstrual phases. CONCLUSION: Levels of immunoreactive endothelin-1 and immunoreactive big endothelin-1 are different in each type of uterine tissue and in each phase of the menstrual cycle. These changes may indicate some role of endothelin-1 in menstruation.

Antibodies↗

Discriminative stimulus properties of the stereoisomers of the phosphodiesterase inhibitor rolipram.

The discriminative stimulus properties of the specific type IV phosphodiesterase inhibitor, rolipram, and its two stereoisomers were assessed using standard two-lever drug discrimination procedures in which responding on the appropriate lever was reinforced on a FR10 schedule. In three separate drug cues based on training rats to discriminate the racemate (0.2 mg/kg, IP), the (-)-isomer (0.1 mg/kg), or the (+)-isomer (2 mg/kg) from vehicle, all forms substituted for one another, differing only in potency. In keeping with published reports, the (-)-isomer was the more potent form, the (+)-isomer being approximately 10 times less potent. Several phosphodiesterase (PDE) inhibitors were found to substitute for the racemate cue, their potencies in the behavioural measure correlating with their potency in displacing [3H]rolipram from its forebrain binding sites in vivo (r = 0.95), suggesting that the discriminative stimulus depends on an action of the drug upon this site. Because rolipram has been reported to possess antidepressant activity, the ability of the tricyclic antidepressant imipramine to substitute for rolipram was investigated; doses of 10 and 20 mg/kg did not substitute. Amphetamine (0.156-1.25 mg/kg) also was inactive. Lisuride gave rise to drug-appropriate responding in 50% of rats only at a dose of 0.078 mg/kg, which severely disrupted responding. It is concluded that the rolipram discriminative stimulus is dependent on the selective PDE inhibitory activity of the drug, and that it does not constitute a cue based on the antidepressant property of rolipram.

Animals↗